key: cord-329944-ywusapij authors: Harbourt, D.; Haddow, A.; Piper, A.; Bloomfield, H.; Kearney, B.; Gibson, K.; Minogue, T. title: Modeling the Stability of Severe Acute Respiratory Syndrome Coronavirus 2 (SARS-CoV-2) on Skin, Currency, and Clothing date: 2020-07-03 journal: nan DOI: 10.1101/2020.07.01.20144253 sha: doc_id: 329944 cord_uid: ywusapij A new coronavirus (SARS-CoV-2) emerged in the winter of 2019 in Wuhan, China, and rapidly spread around the world. The extent and efficiency of SARS-CoV-2 pandemic is far greater than previous coronaviruses that emerged in the 21st Century. Here, we modeled stability of SARS-CoV-2 on skin, paper currency, and clothing to determine if these surfaces may factor in the fomite transmission dynamics of SARS-CoV-2. Skin, currency, and clothing samples were exposed to SARS-CoV-2 under laboratory conditions and incubated at three different temperatures (4C, 22C, and 37C). Stability was evaluated at 0 hours (h), 4 h, 8 h, 24 h, 72 h, 96 h, 7 days, and 14 days post-exposure. SARS-CoV-2 was shown to be stable on skin through the duration of the experiment at 4C (14 days). Virus remained stable on skin for at least 96 h at 22C and for at least 8h at 37C. There were minimal differences between the tested currency samples. The virus remained stable on the $1 U.S.A. Bank Note for at least 96 h at 4C while viable virus was not detected on the $20 U.S.A. Bank Note samples beyond 72 h. The virus remained stable on both Bank Notes for at least 8 h at 22C and 4 h at 37C. Clothing samples were similar in stability to the currency with the virus being detected for at least 96 h at 4C and at least 4 h at 22C. No viable virus was detected on clothing samples at 37C after initial exposure. This study confirms the inverse relationship between virus stability and temperature. Furthermore, virus stability on skin demonstrates the need for continued hand hygiene practices to minimize fomite transmission both in the general population as well as workplaces where close contact is common. 4°C and at least 4 h at 22°C. No viable virus was detected on clothing samples at 37°C after 48 initial exposure. This study confirms the inverse relationship between virus stability and 49 temperature. Furthermore, virus stability on skin demonstrates the need for continued hand 50 hygiene practices to minimize fomite transmission both in the general population as well as 51 workplaces where close contact is common. was not certified by peer review) is the author/funder, who has granted medRxiv a license to display the preprint in perpetuity. The copyright holder for this preprint (which this version posted July 3, 2020. The emergence of SARS-CoV-2 represents the third major outbreak of a new human coronavirus 57 disease over the past twenty years. This novel coronavirus (SARS-CoV-2) that initially emerged 58 in Wuhan, China in late 2019 resulted in a global pandemic that, as of April 22, 2020, has 59 officially resulted in more than 2.5 million cases and 175,000 deaths (1) (2) (3) . The rapid and 60 extensive spread of the virus could be indicative of both aerosol and fomite transmission which 61 has been seen in previous coronavirus outbreaks (4) . Previous studies have shown that SARS-62 CoV-2 is stable at room temperature on stainless steel for approximately 24 hours (h) and on 63 cardboard for up to three days (5) (6) . Additional studies have shown an inverse relationship 64 between surface temperature and stability which is consistent with previous reporting on stability 65 of human coronaviruses (5) . investigations into stability on skin specimens and paper currency (7) . Despite limited evidence, 69 some countries have taken measures to limit the spread of the virus by either burning or 70 disinfecting paper currency or discouraging the use of cash during transactions (8) . It would be 71 expected that the stability of SARS-CoV-2 on currency would be similar to cardboard given that 72 both surfaces are porous, but the effect of ink and toner on the virus remains unknown (6) . 73 Furthermore, it was not known how long SARS-CoV-2 could remain viable on human or animal 74 skin as no similar studies had been performed to date. Handwashing and hand hygiene have been 75 a key part of mitigation efforts, but fomite transmission likely remains a contributing factor to 76 All rights reserved. No reuse allowed without permission. was not certified by peer review) is the author/funder, who has granted medRxiv a license to display the preprint in perpetuity. The copyright holder for this preprint (which this version posted July 3, 2020. . https://doi.org/10.1101/2020.07.01.20144253 doi: medRxiv preprint 4 the speed and extent of the pandemic (9-11). Herein, we model the stability of SARS-CoV-2 77 across animal skin, paper currency, and clothing. was not certified by peer review) is the author/funder, who has granted medRxiv a license to display the preprint in perpetuity. The copyright holder for this preprint (which this version posted July 3, 2020. were disinfected using 5% Microchem Plus™ followed by 70% ETOH between samples. Samples were stored at -80°C prior to virus quantification. incubated at 37⁰C and 5% CO2 for one hour, with rocking approximately every 15 minutes. Following incubation, media-agarose overlay (2mL of a 1:1 mixture of 1.0% agarose and 2X 116 EBME + 10% HI FBS + 2% penicillin, 2% streptomycin + 1% fungizone (2X EBME 117 Complete)) was added to each well. Once overlay solidified, plates were incubated at 37⁰C and 118 5% CO2 for 48 h ± 4 h. Following incubation, a second media agarose overlay containing 4% 119 neutral red in a 1:1 mixture of 1.0% agarose and 2X EBME complete was added to each well. Once overlay solidified, plates were incubated at 37⁰C and 5% CO2 overnight. Following 121 incubation, plaques were counted and the virus yield (plaque forming unit, PFU/mL) for each 122 All rights reserved. No reuse allowed without permission. was not certified by peer review) is the author/funder, who has granted medRxiv a license to display the preprint in perpetuity. The copyright holder for this preprint (which this version posted July 3, 2020. . https://doi.org/10.1101/2020.07.01.20144253 doi: medRxiv preprint 6 sample calculated, with a lower limit of quantification (LLOQ) of 2.0 log10 PFU/mL. The limit 123 of detection (LOD) for virus isolation attempts was 0.1 log10 PFU/mL. The mixed model procedure was used to allow for a heterogeneous variance structure (11). was not certified by peer review) is the author/funder, who has granted medRxiv a license to display the preprint in perpetuity. The copyright holder for this preprint (which this version posted July 3, 2020. was not certified by peer review) is the author/funder, who has granted medRxiv a license to display the preprint in perpetuity. The copyright holder for this preprint (which this version posted July 3, 2020. (Fig 2) . Virus was not isolated from the $1 U.S.A. Bank Note or clothing beyond 8 h at 185 room temperature. Viable virus was detected on the $20 U.S.A. Bank Note at 24 h but all 186 subsequent samples were negative for infectious virus (Fig 1 and Fig 2) . Approximately 3 log10 188 h but all other tested samples were below the LOD. was not certified by peer review) is the author/funder, who has granted medRxiv a license to display the preprint in perpetuity. The copyright holder for this preprint (which this version posted July 3, 2020. . https://doi.org/10.1101/2020.07.01.20144253 doi: medRxiv preprint 9 190 SARS-CoV-2 remained viable at 37°C on skin samples for up to 8 h (Fig 1 and Fig 2) . No 191 viable, infectious virus was detected on clothing samples after four hours at 37°C (Fig 2) . There 192 were minor differences in stability at 37°C between the $1 U.S.A. Bank Note and the $20 U.S.A. Bank Note, but those differences were not statistically significant. There was a discernable (Table 1 ). Significant differences in virus stability 199 were noted when each temperature condition was compared (Fig S1) . Significant differences 200 were observed in virus stability between the skin samples and all other tested surfaces (Fig S2) . was not certified by peer review) is the author/funder, who has granted medRxiv a license to display the preprint in perpetuity. The copyright holder for this preprint (which this version posted July 3, 2020. was not certified by peer review) is the author/funder, who has granted medRxiv a license to display the preprint in perpetuity. The copyright holder for this preprint (which this version posted July 3, 2020. It is important to note the limitations of this study when analyzing the results. The variability in 252 the decay curves between earlier studies and our results could have been due to differences in 253 All rights reserved. No reuse allowed without permission. was not certified by peer review) is the author/funder, who has granted medRxiv a license to display the preprint in perpetuity. The copyright holder for this preprint (which this version posted July 3, 2020. . https://doi.org/10.1101/2020.07.01.20144253 doi: medRxiv preprint 12 exposure doses (4, 6, 7) . In addition, inherent variability with a study where virus is recovered 254 from a tested surface can potentially confound some of the results, particularly at virus 255 concentrations close to the limit of detection (LOD) or lower limit of quantification (LLOQ) 256 (Fig. 1) . The results in this study demonstrate the continued inverse relationship between virus stability 259 and temperature which is seen both in the laboratory and in the field when evaluating different was not certified by peer review) is the author/funder, who has granted medRxiv a license to display the preprint in perpetuity. The copyright holder for this preprint (which this version posted July 3, 2020. was not certified by peer review) is the author/funder, who has granted medRxiv a license to display the preprint in perpetuity. The copyright holder for this preprint (which this version posted July 3, 2020. . https://doi.org/10.1101/2020.07.01.20144253 doi: medRxiv preprint Coronavirus disease (COVID-2019) situation reports. 2020. (Accessed 22 nd of April A Novel Coronavirus from Patients with Pneumonia in China A novel coronavirus outbreak of global health 308 concern Transmission 311 of SARS and MERS coronaviruses and influenza virus in healthcare settings: the possible 312 role of dry surface contamination Persistence of coronaviruses on inanimate 315 surfaces and their inactivation with biocidal agents Aerosol and Surface Stability of SARS-CoV-2 as Compared with SARS No reuse allowed without permission. was not certified by peer review) is the author/funder, who has granted medRxiv a license to display the preprint in perpetuity. The copyright holder for this preprint Stability of SARS-CoV-2 in different environmental conditions Business Insider Stability and 332 inactivation of SARS coronavirus Human Coronaviruses: Insights into Environmental Resistance and Its Influence on the Development of New Antiseptic 336 Strategies. Viruses Influence in Heterogeneous Variance Model User's Guide: The Mixed Procedure R: A language and environment for statistical computing. R 343 Foundation for Statistical Computing No reuse allowed without permission. was not certified by peer review) is the author/funder, who has granted medRxiv a license to display the preprint in perpetuity. The copyright holder for this preprint Effects of air temperature 347 and relative humidity on coronavirus survival on surfaces Face touching: a frequent habit that has implications 351 for hand hygiene Skin xenotransplantation: 355 Historical review and clinical potential Disinfection of transboundary 358 animal disease viruses on surfaces used in pork packing plants Antimicrobial and Antioxidative Strategies to Reduce 362 Pathogens and Extend the Shelf Life of Fresh Red Meats SARS-CoV-2 Viral Load in Upper Respiratory Specimens of Infected Patients The authors are indebted to Mr. Brian Sauerbry who assisted with material procurement, Dr. Pamela Glass for technical assistance and Ms. Sarah Norris for assistance with the statistical All rights reserved. No reuse allowed without permission.was not certified by peer review) is the author/funder, who has granted medRxiv a license to display the preprint in perpetuity.The copyright holder for this preprint (which this version posted July 3, 2020. was not certified by peer review) is the author/funder, who has granted medRxiv a license to display the preprint in perpetuity.The copyright holder for this preprint (which this version posted July 3, 2020. was not certified by peer review) is the author/funder, who has granted medRxiv a license to display the preprint in perpetuity.The copyright holder for this preprint (which this version posted July 3, 2020. . https://doi.org/10.1101/2020.07.01.20144253 doi: medRxiv preprint